Bryostatin activates HIV-1 latent expression in human astrocytes through a PKC and NF-ĸB-dependent mechanism.
Bryostatin activates HIV-1 latent expression in human astrocytes through a PKC and NF-ĸB-dependent mechanism.
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Bryostatin通过PKC和NF-ĸB依赖性机制激活人类星形胶质细胞中的HIV-1潜在表达。
DOI：
10.1038/srep12442
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发表时间：
2015-07-22
影响因子：
4.6
通讯作者：
Muñoz-Fernández MÁ
中科院分区：
文献类型：
--
作者：
Díaz L;Martínez-Bonet M;Sánchez J;Fernández-Pineda A;Jiménez JL;Muñoz E;Moreno S;Álvarez S;Muñoz-Fernández MÁ
多项研究表明，HIV-1 患者可能会形成病毒库，从而阻碍根除；这些储存库包括中枢神经系统（CNS）。尽管接受强效抗逆转录病毒药物治疗的患者体内病毒载量无法检测到，但目前的治疗无法清除这些潜伏病毒库中的病毒。为了扩大现有抗逆转录病毒药物的抑制范围和有效性，研究了苔藓抑素作为潜在 HIV-1 激活剂的潜力。我们使用原代星形胶质细胞、NHA 细胞和星形细胞瘤细胞 U-87。单独用苔藓抑素或与不同抑制剂联合处理感染 HIV-1NL4.3 的细胞。 HIV-1 的产生通过 ELISA 进行定量。转录活性通过使用lipofectin的荧光素酶报告基因测定来测量。我们进行了 LTR 启动子与活性 NF-κB 成员 p65/relA 的共转染实验。为了证实 NF-κB 的作用，进行了蛋白质印迹和共聚焦显微镜检查。苔藓抑素通过激活蛋白激酶 C (PKC)-α 和 δ 来重新激活 NHA 和 U87 细胞中的潜伏病毒感染，因为 PKC 抑制剂 rottlerin 和 GF109203X 消除了苔藓抑素的作用。没有发现细胞增殖的改变。此外，苔藓抑素通过激活转录因子 NF-κB 强烈刺激 LTR 转录。苔藓抑素可能是治疗 HIV-1 脑部感染的有益辅助药物。
Multiple studies have shown that HIV-1 patients may develop virus reservoirs that impede eradication; these reservoirs include the central nervous system (CNS). Despite an undetectable viral load in patients treated with potent antiretrovirals, current therapy is unable to purge the virus from these latent reservoirs. To broaden the inhibitory range and effectiveness of current antiretrovirals, the potential of bryostatin was investigated as a latent HIV-1 activator. We used primary astrocytes, NHA cells, and astrocytoma cells U-87. Infected cells with HIV-1NL4.3 were treated with bryostatin alone or in combination with different inhibitors. HIV-1 production was quantified by using ELISA. Transcriptional activity was measured using luciferase reporter gene assays by using lipofectin. We performed cotransfection experiments of the LTR promoter with the active NF-κB member p65/relA. To confirm the NF-κB role, Western blot and confocal microscopy were performed. Bryostatin reactivates latent viral infection in the NHA and U87 cells via activation of protein kinase C (PKC)-alpha and -delta, because the PKC inhibitors rottlerin and GF109203X abrogated the bryostatin effect. No alteration in cell proliferation was found. Moreover, bryostatin strongly stimulated LTR transcription by activating the transcription factor NF-κB. Bryostatin could be a beneficial adjunct to the treatment of HIV-1 brain infection.